Aptamer Cocktail to Detect Multiple Species of Mycoplasma in Cell Culture

Int J Mol Sci. 2020 May 27;21(11):3784. doi: 10.3390/ijms21113784.

Abstract

Mycoplasma contamination of cell line cultures is a common, yet often undetected problem in research laboratories. Many of the existing techniques to detect mycoplasma contamination of cultured cells are time-consuming, expensive, and have significant drawbacks. Here, we describe a mycoplasma detection system that is useful for detecting multiple species of mycoplasma in infected cell lines. The system contains three dye-labeled detection aptamers that can specifically bind to mycoplasma-infected cells and a dye-labeled control aptamer that minimally binds to cells. With this system, mycoplasma-contaminated cells can be detected within 30 min by using a flow cytometer, fluorescence microscope, or microplate reader. Further, this system may be used to detect mycoplasma-contaminated culture medium. This study presents an novel mycoplasma detection model that is simple, rapid, inexpensive, and sensitive.

Keywords: DNA aptamer; cell culture; mycoplasma; rapid detection.

MeSH terms

  • Aptamers, Nucleotide / metabolism*
  • Binding, Competitive
  • Cell Culture Techniques*
  • Cell Line, Tumor
  • Culture Media
  • DNA Contamination
  • Flow Cytometry
  • Humans
  • Mycoplasma / genetics
  • Mycoplasma / isolation & purification*

Substances

  • Aptamers, Nucleotide
  • Culture Media